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A thermophilic fungus, Talaromyces thermophilus that produces a novel thermotolerant extra-cellular β-glucosidase (Bgl.tls), was isolated from Tunisian soil samples. The enzyme was purified from the culture filtrates of T. thermophilus grown on lactose using gel filtration, ion exchange chromatography and FPLC. The monomeric enzyme had a molecular mass of 116.0 kDa and a high specific activity of 1429 UI/mg. Bgl.tls exhibited optimal activity at pH 5.0 and 65 °C. It was also stable over a wide range of pH (4.0-10.0) and stable at 50 °C for 34 h. Bgl.tls retained about 80% of its initial activity after 1.0 hours of preincubation at 60 °C. The Km and Vmax values recorded for pNPG were 0.25 mM and 228.7 µmol min(-1), respectively. Bgl.tls was activated by Mn(2+), Mg(2+), Ca(2+) and Co(2+) but obviously inhibited by Fe(2+) and Cu(2+). It was able to hydrolyze a variety of aryl / alkyl -β-glucosides and disaccharides as well as (1 → 6) and (1 → 4)-β-glucosidic linkages and α-glycosidic substrates, thus providing evidence for its broad substrate specificity. The enzyme also displayed high hydrolytic and transglycosylation activities. Overall, this study is the first report on the purification and physicochemical properties of a β-glucosidase secreted by T. thermophilus. The cello-oligosaccharides synthesized by this enzyme within 2 h were mainly cellotriose, cellotetraose and cellopentaose identified by HPLC and ESI-MS techniques., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The effect of high hydrostatic pressure treatment (with pressures of up to 700 MPa) on Talaromyces macrosporus ascospores was investigated. At 20 degrees C, pressures of > or = 200 MPa induced the activation and germination of dormant ascospores, as indicated by increased colony counts for ascospore suspensions after pressure treatment and the appearance of germination vesicles and tubes. Pressures of > 400 MPa additionally sensitized the ascospores to subsequent heat treatment. At pressures of > 500 MPa, activation occurred in a few minutes but was followed by inactivation with longer exposure. However, even with the most extreme pressure treatment, a fraction of the ascospore population appeared to resist both activation and inactivation, and the maximal achievable reduction of ascospores was on the order of 3.0 log10 units. Pressure-induced ascospore activation at 400 MPa was temperature dependent, with minimum activation at 30 to 50 degrees C and > or = 10-fold higher activation levels at 10 to 20 degrees C and at 60 degrees C, but it was not particularly pH dependent over a pH range of 3.0 to 6.0. Pressure inactivation at 600 MPa, in contrast, was pH dependent, with the inactivation level being 10-fold higher at pH 6.0 than at pH 3.0. Observation of pressure-treated and subsequently dried spores with the use of light and scanning electron microscopy revealed a collapse of the spore structure, indicating a loss of the spore wall barrier properties. Finally, pressure treatment sensitized T. macrosporus ascospores to cell wall lytic enzymes.